Time resolution of binding and membrane insertion of a mitochondrial signal peptide: Correlation with structural changes and evidence for cooperativity

被引:37
作者
Golding, C
Senior, S
Wilson, MT
OShea, P
机构
[1] UNIV ESSEX,DEPT BIOL & CHEM SCI,COLCHESTER CO4 3SQ,ESSEX,ENGLAND
[2] APPL PHOTOPHYS LTD,LEATHERHEAD KT22 7PB,SURREY,ENGLAND
关键词
D O I
10.1021/bi960905i
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Utilizing a recently developed novel fluorescence technique [Wall et al. (1995) Mol. Membr. Biol. 12, 183-192], it is shown that the interactions of p25, the leader peptide of subunit IV of cytochrome c oxidase, with phospholipid membranes can be identified in real time. p25 is observed to bind following stopped-flow mixing of the peptide with phospholipid membranes with rate constants up to about 700 s(-1) and then insert into the membrane with rate constants on the order of 0.4 s(-1). Comparison of these processes with similarly time-resolved experiments performed with a stopped-flow CD spectrometer revealed that p25 does not become alpha-helical upon binding to the membrane. Following membrane insertion, however, p25 was observed to adopt an alpha-helical configuration. The temperature dependency of these processes was then found to yield activation energies for the respective components of the p25-membrane interaction.
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页码:10931 / 10937
页数:7
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